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Main sol. 311b

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Internal Solution-ID: S578

Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 0.50 g 0.498 9.301
MgSO4 x 7 H2O 0.50 g 0.498 2.019
CaCl2 x 2 H2O 0.25 g 0.249 1.692
NaCl 25.00 g 24.876 425.661
FeSO4 x 7 H2O
(0.1% w/v in 0.1 N H2SO4) 		2.00 ml 0.002 0.007
Trace element solution SL-10 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
Yeast extract 2.00 g 1.99 -
Casitone 2.00 g 1.99 -
Betaine x H2O 6.70 g 6.667 56.909
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
K2HPO4 0.35 g 0.348 1.999
KH2PO4 0.23 g 0.229 1.682
Na2CO3 1.50 g 1.493 14.082
Wolin's vitamin solution (10x) 1.00 ml - -
L-Cysteine HCl x H2O 0.30 g 0.299 1.7
Na2S x 9 H2O 0.30 g 0.299 1.243
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except phosphates, carbonate, vitamins, cysteine and sulfide) and sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense under same gas atmosphere in anoxic Hungate-type tubes or serum vials and autoclave. Add phosphates, vitamins (sterilized by filtration), cysteine and sulfide to the medium after autoclaving from sterile stock solutions prepared under 100% N2 gas and carbonate from a sterile stock solution prepared under 80% N2 and 20% CO2 gas mixture. Adjust pH of completed medium to pH 7.5 - 7.7.
311b SPOROMUSA MEDIUM (MARINE)